At the human GATA2 locus, disruption of two distinct cis-elements (+9.5 and -77) instigate immunodeficiency,myelodysplastic syndrome and/or acute myeloid leukemia. To define essential GATA-2 target genes, we implemented a multi-omics prioritization strategy based on sequence and molecular criteria. Among others, this analysis revealed GATA-2 regulation of Sterile Alpha Motif(SAM) Domain 14 (Samd14) expression. Mechanistic studies established that Samd14 (and the GATA-2-occupied Samd14 enhancer (Samd14-Enh)) promotes hematopoietic stem/progenitor cell (HSPC) function, facilitates regeneration in adult hematopoiesis, and is required for survival in severe anemia. Samd14-Enh promotes cell signaling through the oncogenic SCF/c-Kit pathway, conferring survival. The key mechanisms whereby Samd14 promotes SCF/c-Kit signaling and hematopoietic progenitor function are not understood. Anemia-activated Gata2 and Samd14 transcription is enhancer-dependent, establishing a foundation for defining a GATA-2 and anemia-activated gene network in regeneration, involving many intriguing and unstudied network constituents. Utilizing COBRE-sponsored facilities at UNMC and collaborations with COBRE program leaders, these aims will rigorously establish how Samd14 promotes SCF/c-Kit signaling (Aim 1), the hematopoietic function of a new SAM-domain containing gene (Aim 2), and the function of Samd14 in hematopoietic contexts (Aim 3). The proposed project is highly relevant to the overall mission of the ?Nebraska Center for Molecular Target Discovery and Development?. We will define completely novel molecular mechanisms regulating an integral, oncogenic cell signaling pathway (SCF/c-Kit) with the goal of identifying therapeutic targets. The combination of Project Leader collaborations, resources provided by COBRE Cores, and the support of scientific/career mentorship will greatly enhance the impact of scientific discoveries and success of competing for R01-level funding.